JP2023106431A - Compounds for use in prevention and/or treatment of non-alcoholic fat liver disease and non-alcoholic steatohepatitis - Google Patents

Abstract

To provide pharmaceutical compositions for use in the prevention and/or treatment of non-alcoholic steatohepatitis (NASH).SOLUTION: The present invention discloses at least one compound selected from the following compounds 4a, 5a and the like.SELECTED DRAWING: None

Description

The present invention provides prevention and/or treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), as well as prevention and/or treatment of all episodes or symptoms associated with these liver diseases. Of interest is a family of compounds for use in

Nonalcoholic fatty liver disease (NAFLD) and its severe development, nonalcoholic steatohepatitis (NASH), is an emerging therapeutic area. NAFLD is characterized by an abnormal accumulation of fat in the liver as lipid droplets within the liver parenchyma and affects individuals who consume less alcohol (1,2). It is the most frequent liver disorder worldwide, affecting 6-35% of the general population and a leading cause of liver transplantation after progression to cirrhosis and hepatocellular carcinoma (3). is one of

The actual cause of NAFLD remains unknown. However, insulin resistance, among other diseases, is believed to play a role in the disease process. The actual reasons and mechanisms by which the disease progresses from one stage to the next are not fully understood.

The development of the disease has been explained by the "two-hit hypothesis", in which the first blow is represented by lipid accumulation in hepatocytes, followed by the second blow oxidative stress and inflammation leading to NASH ( 1). The early metabolic stress generated by lipid accumulation in hepatocytes triggers multiple cellular stress pathways, including endoplasmic reticulum stress, mitochondrial dysfunction, oxidative stress (production of reactive oxygen species), apoptosis, and even necrosis. . The hepatocellular injury produced results in the release of signals that recruit and activate various immune cells leading to an inflammatory response. Collectively, these events lead to activation of hepatic stellate cells, including increased collagen deposition that results in fibrosis, which may eventually progress to cirrhosis and hepatocellular carcinoma (4). Other external inputs from behavioral habits (diet/lifestyle/exercise) to microbiota molecules can contribute to disease development (5).

To date, many therapeutic strategies have been proposed and drug candidates tested. However, none of them have yet been approved for the treatment of NASH (6-8). Different targets and strategies have been explored in searching for effective therapies to treat NAFDL and NASH. This includes:
• Insulin sensitizers: PPAR agonists, incretin analogs (GLP-1 receptor agonists), DPP-4 inhibitors, SGLT2 inhibitors, ACE inhibitors, and angiotensin II receptor antagonists (antihypertensives).
Bile acid regulators: Farnesoid X receptor agonists (negatively modulates bile acid synthesis and reduces hepatic lipogenesis and steatosis)
De novo lipogenesis inhibitors: stearoyl-CoA desaturase and acetyl-CoA carboxylase inhibitors Lipid-lowering drugs: statins, fibrates, lipase inhibitors Antioxidants: vitamin E, cysteamine Anti-inflammatory drugs: TNF-α inhibitors Immunity Modulating components: IκB inhibitors, inflammatory chemokines antagonists (CCR2/CCR5 inhibitors), VAP1 inhibitors Anti-apoptotic agents: caspase inhibitors, ASK1 inhibitors Gut microbiome modulators: antibiotics, anti-LPS IgG-rich extract stool microbial transplantation/antifibrotic agent: galectin-3 inhibitor, LOXL2 blocker

Galectin-3 is considered a target for fibrosis, but is also a marker for hepatitis, as measured in the examples included in this disclosure. As noted above, none of these approaches have yet resulted in approved compounds for the treatment of NASH, thus there is no need to develop alternatives to these approaches for the treatment and prevention of said disease. Obvious.

The present invention provides compounds for use in methods of reducing liver fat content associated with non-alcoholic fatty liver disease or steatohepatitis. In particular, the subject of the present disclosure is compounds and methods for the prevention and/or treatment of non-alcoholic fatty liver disease (NAFLD) and its severe developmental stage, non-alcoholic steatohepatitis (NASH). to provide.

NAFLD is thought to encompass a spectrum of disease activity. This spectrum begins with fat accumulation in the liver (fatty liver), as explained. Although the liver can remain fatty in the absence of disturbance of liver function, different mechanisms and possible hepatic aggression proceed, resulting in the development of non-alcoholic steatohepatitis (NASH). In this condition, steatosis is accompanied by inflammation and fibrosis (steatohepatitis).

Thus, the progression of this disease from NAFLD to NASH includes a variety of symptoms and/or associated pathologies directly linked to the disorder.

With respect to the early stages, the symptoms and/or associated pathologies trigger multiple cellular stress pathways, including endoplasmic reticulum stress, mitochondrial dysfunction, oxidative stress (production of reactive oxygen species), apoptosis, and even necrosis. Refers to resistance and lipid accumulation in hepatocytes.

In the second stage, the hepatocellular injury produced leads to symptoms of NAFLD associated with inflammation.

Collectively, these events result in activation of hepatocytes, including increased fibrosis, increased collagen deposition leading to cirrhosis and hepatocellular carcinoma as associated pathologies of these disorders.

Accordingly, the problem to be solved by the present invention is to provide compounds of formula I for use in the prevention and/or treatment of NAFLD and NASH symptoms and/or related pathologies at all stages of their development. yes; preferably the associated condition is independently selected from insulin resistance, intrahepatocyte lipid accumulation, mitochondrial dysfunction, oxidative stress, apoptosis, necrosis, inflammation, or fibrosis; The underlying pathology is cirrhosis or hepatocellular carcinoma.

The compounds of formula I disclosed in the present invention and their synthesis were disclosed in International Patent Application PCT/EP2012/055570 filed on March 28, 2012. According to PCT/EP2012/055570 said compounds are useful for the treatment of metabolic syndrome. Applicants have now surprisingly found that said compounds and their synthetic intermediates are further useful for the prevention and/or treatment of NAFLD or NASH, as well as the development and/or symptoms associated therewith. I have discovered that

Accordingly, the present invention provides general With respect to compounds of formula I, and pharmaceutically or food acceptable salts thereof:

where independently
R ₁ can be selected from: linear or cyclic, mono- or dialkylamines; OR ₉ , aminoalkyl alcohols or aminoalkyl ethers;
R ₂ can be selected from a benzene ring or a heterocycle;
R ₃ can be selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; or a benzyl group;
R ₄ can be selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; a hydroxy or alkoxy radical; or a halogen;
_R9 is an alkyl group.

In one embodiment, the compounds described herein are for use in the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), and/or symptoms and/or medical conditions associated therewith. , wherein the relevant condition is independently selected from insulin resistance, lipid accumulation in hepatocytes, mitochondrial dysfunction, oxidative stress, apoptosis, necrosis, inflammation or fibrosis. and associated conditions are cirrhosis or hepatocellular carcinoma.

Preferred compounds of general formula I for use according to the invention are independently
When R ₁ is a linear alkylamine: NH(CH ₂ ) _n —NH ₂ , NH(CH ₂ ) _n —N(CH ₃ ) ₂ (where n is a value of 0 to 4); NH—N =CH-phenyl-R ₇ ;
and R ₁ is selected from the following when it is a cyclic amine:

When an aminoalkyl alcohol group, R ₁ is HNCH ₂ CH ₂ OH; and when an aminoalkyl ether group, R ₁ is HNCH ₂ CH ₂ OCH ₃ ;
when OR ₉ , R ₉ , R ₁ is a C _1-4 alkyl group;
When it is a benzene substituted ring, _R2 is selected from:

and heterocycle, then

when it is a hydrocarbon radical selected from linear alkyl of 1 to 5 carbons, R ₃ is methyl;
when it is a hydrocarbon radical selected from linear alkyl of 1 to 5 carbons, R ₄ is methyl;
when an alkoxy radical, R ₄ is a methoxy radical; and
when halogen, R ₄ is fluorine;
_alkoxy ; halogen; hydroxy; or halogen-alkyl;
_R7 can be selected from H or _NO2 ;
_R8 can be selected from H; hydroxy; alkoxy.

The term "pharmaceutically acceptable salt" refers to any salt that is capable (directly or indirectly) of the compounds described herein upon administration to a patient.

The term "food acceptable salt" refers to any salt of the described product that can be added to a functional food additive or dietary supplement.

Such salts are preferably acid addition salts with physiologically acceptable organic or inorganic acids. Examples of acid addition salts include mineral acid addition salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphoric acid and, for example, acetate, trifluoroacetic acid, maleate. , fumarate, citrate, oxalate, succinate, tartarate, malate, mandelate, methanesulfonate, and organic acid addition salts such as p-toluenesulfonic acid. Examples of alkaline addition salts include inorganic salts such as sodium, potassium, calcium, and ammonium salts, and organic salts such as ethylenediamine, ethanolamine, N,N-dialkylethanolamine, triethanolamine, and basic amino acid salts. Alkaline salts are given.

In a preferred embodiment of the invention the salt is the hydrochloride.

Accordingly, the present invention provides compounds of Formula I, and pharmaceutical compounds thereof, as described herein above, for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or pathologies associated therewith. Or refers to acceptable salt for food.

The present invention also relates to a compound of formula I and its pharmaceutical or food use for the manufacture of a medicament for the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or pathologies associated therewith. It refers to the use of acceptable salts.

Additionally, the present invention refers to methods of preventing and/or treating a subject suffering from or suffering from symptoms associated with NAFLD or NASH and/or suffering from associated medical conditions, which methods are disclosed herein. administering to said subject an effective amount of a compound of Formula I, and a pharmaceutically or food acceptable salt thereof.

Preferred compounds for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or pathologies associated therewith, as described in the present invention are compounds of Formula I, wherein R ₅ is selectable from: H, methoxy; chlorine, OH, or trifluoromethyl, preferably when _R5 is H, _{R8 is OH, and when R5 is} OH, _R8 is H or _OCH3 .

A preferred compound for use according to the invention is a compound of formula I, wherein _R9 is methyl.

Preferred compounds for use according to the invention are compounds of formula I, wherein R ₂ is:

Preferred compounds for use according to the invention are compounds of formula I, wherein _R3 is methyl or benzyl.

Preferred compounds for use according to the invention are compounds of formula I, wherein _R4 is methyl, methoxy, or fluorine.

Preferred compounds according to the present disclosure are compounds of formula I(I) or a pharmaceutically acceptable is a salt,

During the ceremony,
—R ₁ is OR ₉ ; NH(CH ₂ ) _n —NH ₂ , NH(CH ₂ ) _n —N(CH ₃ ) ₂ (where n is a value of 0 to 4); HNCH ₂ CH ₂ OH; ₂ CH ₂ OCH ₃ ; NH—N═CH-phenyl-R ₇ ; or a cyclic amine;
- _R2 is selected from

—R ₃ can be selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; or a benzyl group;
—R ₄ can be selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; a hydroxy or alkoxy radical; or a halogen;
-R ₇ is H or p-NO ₂ and -R ₅ is selected from H; alkoxy; halogen; hydroxy; or halogen-alkyl;
—R ₈ is selected from H; hydroxy; alkoxy;
-R ₉ is methyl.

Additionally, preferred compounds of the present disclosure are compounds of formula I(I) or pharmaceutical compounds thereof for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or medical conditions associated therewith. is a legally acceptable salt,

During the ceremony,
—R ₁ is NH(CH ₂ ) _n —NH ₂ , NH(CH ₂ ) _n —N(CH ₃ ) ₂ (where n is a value of 0 to 4); HNCH ₂ CH ₂ OH; HNCH ₂ CH ₂ NH—N═CH-phenyl-R ₇ ; or a _cyclic amine selected from:

- _R2 is selected from

-R ₃ is selected from H, methyl, or benzyl;
-R ₄ is selected from H, methyl, methoxy, or fluorine;
-R ₇ is H or p- _NO2 ;
—R ₈ is H, OH, or methoxy; and
—R ₅ is H, OH, or methoxy.

In preferred embodiments, the pharmaceutically acceptable salt is the hydrochloride salt.

Additionally preferred compounds for use in accordance with the present invention are compounds having formula II or III,

where independently
R ₁ is; OH, OCH ₃ , NH—(CH ₂ ) _n —N(CH 3 ) ₂ , NH—(CH ₂ ) _n —NH ₂ where n is a value of 0 to ₃ ; NH(CH ₂ ) ₂ -OH; NH-(CH ₂ ) ₂ -OCH ₃ , or NH-N=CH-phenyl-R ₇ ;
R ₅ can be selected from OCH ₃ or H;
R ₇ can be selected from H or p-NO ₂ .

More particularly preferred compounds for use according to the invention are those in which R ₅ is selected from H or p-OCH ₃ when R ₁ is an OH group.

Preferred further compounds for use according to the invention are from formula _{1a ,} wherein R ₁ is an OH group and R ₅ is p-OCH ₃ ; A compound having Formula II selected from certain Formulas 1b.

Further preferred compounds for use according to the invention are those in which R ₅ is selected from H or p-OCH ₃ when R ₁ is an OCH ₃ group.

Preferred compounds for use according to the invention are: Formula 2a, wherein R ₁ is the OCH ₃ group and R ₅ is p-OCH ₃ ; R ₁ is the OCH ₃ group and R ₅ is H or Formula 3a wherein R ₁ is an OCH ₃ group and R ₅ is p-OCH ₃ , or wherein R ₁ is an OCH ₃ group and R ₅ is H A compound having formula III selected from 3b.

Further preferred compounds for use according to the present invention are compounds wherein R ₅ is p-OCH ₃ when R ₁ is NH—(CH ₂ ) _n —NH ₂ and n=2 or a value of 3. It is a compound.

Preferred compounds for use according to the invention are from Formula 4a, wherein R ₁ is NH(CH ₂ ) ₂ NH ₂ and R ₅ is p-OCH ₃ ; or R ₁ is NH(CH ₂ ) ₃ NH ₂ and R ₅ is p-OCH ₃ . Formula III selected from Formula 5a.

A more preferred compound for use according to the present invention is 4a, wherein when R ₁ is a NH—(CH ₂ ) _n —NH ₂ group, n=2, R ₅ is p-OCH ₃ be.

A more preferred compound for use according to the present invention is 5a, wherein when R ₁ is a NH—(CH ₂ ) _n —NH ₂ group, n=3, R ₅ is p-OCH ₃ be.

More preferred compounds for use in accordance with the present invention are compounds having the formula III, wherein when R ₁ is a NH—(CH ₂ )n—NH ₂ group, a value of n=0, R ₅ is H or p- _OCH3 .

Compounds for use according to the present invention are selected from formula 6a wherein R ₁ is NHNH ₂ and R ₅ is p-OCH ₃ ; or formula 6b wherein R ₁ is NHNH ₂ and R ₅ is H further includes a compound having formula III.

More particularly preferred compounds for use according to the present invention are compounds of Formula III when R ₁ is a NH-N=CH-phenyl group, R ₅ is p-OCH ₃ and R ₁ is p-NO Compounds in which R ₅ is H when it is a NH-N=CH-phenyl group substituted by _two groups.

Compounds for use according to the present invention are from formula 7a, wherein R ₁ is a NH-N=CH-phenyl group and R ₅ is p-OCH ₃ ; or R ₁ is NH-N=CH-phenyl-p Further included are compounds having Formula III, selected from Formula 7b, wherein R ₅ is H and is a —NO ₂ group.

Further preferred compounds for use in accordance with the present invention are compounds having the formula III, wherein R ₁ is a NH—(CH ₂ ) _n —N(CH ₃ ) ₂ group with a value of n=2 , R ₅ is p-OCH ₃ .

Further preferred compounds for use in accordance with the present invention are compounds having formula III, wherein R ₁ is a cyclic amine selected from

and R ₅ is p-OCH ₃ .

Further preferred compounds for use in accordance with the present invention are compounds having the formula III, where R ₁ is NH—(CH ₂ ) _n —N(CH ₃ ) ₂ , where n=4 , R ₅ is p-OCH ₃ .

Further preferred compounds for use in accordance with the present invention are compounds having Formula III wherein R ₅ is p-OCH ₃ when R ₁ is NH—(CH ₂ ) ₂ —OH.

Further preferred compounds for use in accordance with the present invention are compounds having formula III wherein when R ₁ is NH—(CH ₂ ) ₂ —OCH ₃ , R ₅ is p-OCH ₃ .

Further preferred compounds for use in accordance with the present invention are compounds having the formula III, where R ₁ is NH—(CH ₂ ) _n —N(CH ₃ ) ₂ , where n=2 , R ₅ is chlorine. Further preferred compounds for use in accordance with the present invention are compounds having the formula III, where R ₁ is NH—(CH ₂ ) _n —N(CH ₃ ) ₂ , where n=2 , _R5 is OH.

Further preferred compounds for use in accordance with the present invention are compounds having formula III, wherein R ₁ is NH—(CH ₂ )n—N(CH ₃ ) ₂ , where n=2 , R ₅ is trifluoromethyl.

Preferred embodiments are for use in the prevention and/or treatment of nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH), and/or symptoms associated therewith and/or medical conditions associated therewith. or a pharmaceutical salt thereof, or a pharmaceutical composition comprising an effective amount of said compound, wherein the relevant condition is insulin resistance, lipids in hepatocytes is independently selected from accumulation, mitochondrial dysfunction, oxidative stress, apoptosis, necrosis, inflammation, or fibrosis; and the associated condition is cirrhosis or hepatocellular carcinoma.

A preferred embodiment is a compound of formula I, II, or III, or a pharmaceutical salt thereof, or a medicament comprising said compound, for use in the prevention and/or treatment of non-alcoholic fatty liver disease (NAFLD) Refers to composition.

A preferred embodiment is a compound of formula I, II, or III, or a pharmaceutical salt thereof, or a pharmaceutical composition comprising said compound, for use in the prevention and/or treatment of non-alcoholic steatohepatitis (NASH) point to things

A preferred embodiment is a compound of formula I, II, or III, or a pharmaceutical salt thereof, or a pharmaceutical composition comprising said compound.

A preferred embodiment is a compound of formula I, II, or III, or a pharmaceutical salt thereof, for use in the prevention and/or treatment of insulin resistance, intrahepatocyte lipid accumulation, hepatitis or liver fibrosis. , or a pharmaceutical composition comprising said compound.

A preferred embodiment refers to a compound of Formula I, II, or III, or a pharmaceutical salt thereof, or a pharmaceutical composition comprising said compound, for use in the prevention and/or treatment of cirrhosis or hepatocellular carcinoma. .

The present invention also optionally includes any inactive ingredients, carriers, excipients, etc. for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or pathologies associated therewith. , at least one of the above compounds represented by general formulas I, II and III, and pharmaceutically or food acceptable or acceptable salts thereof and combinations thereof, pharmaceutical compositions, functional food additives products, or dietary supplements.

The present invention also provides compounds encompassed by general formula I, as previously disclosed, for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or pathologies associated therewith. either, and pharmaceutical salts thereof, or pharmaceutical compositions containing them.

Additionally, the present invention is directed to the prevention and/or treatment of nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH), and/or symptoms and/or medical conditions associated therewith. A pharmaceutical composition comprising an effective amount of at least one compound of formula I as described herein for use.

An effective amount for purposes of the present invention is an amount that provides either subjective relief of symptoms or an objectively identifiable improvement as noted by a clinician or qualified observer. be.

The present invention is also for use in the prevention and/or treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), and/or symptoms and/or medical conditions associated therewith. of a pharmaceutical composition comprising an effective amount of at least one compound of Formula I, and a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient:

where independently
R ₁ is selected from linear or cyclic mono- or dialkylamines; OR ₉ , aminoalkyl alcohols, or aminoalkyl ethers;
R ₂ is selected from benzene or heterocycle;
R ₃ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; or a benzyl group;
R ₄ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; a hydroxy or alkoxy radical; or halogen;
and _R9 are alkyl groups.

Additionally disclosed are pharmaceutical compositions for use as described above comprising an effective amount of at least one compound of Formula I and at least one pharmaceutically acceptable excipient, wherein: —R ₁ is NH—(CH ₂ ) _n —NH ₂ , NH—(CH ₂ ) _n —N(CH ₃ ) ₂ (where n is a value of 0 to 4); HNCH ₂ CH ₂ OH; HNCH ₂ CH ₂ OCH ₃ ; NH—N═CH-phenyl-R ₇ ; or a cyclic amine selected from:

- _R2 is selected from

-R ₃ is selected from H, methyl, or benzyl;
-R ₄ is selected from H, methyl, methoxy, or fluorine;
-R ₇ is H or p-NO ₂ and -R ₈ is H, OH, or methoxy; and
—R ₅ is H, OH, or methoxy.

Additionally, the present invention also provides compounds according to the previously disclosed general formulas I, II or III for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or pathologies associated therewith. Any of the included compounds, and pharmaceutical salts thereof, as well as second active compounds are included. Said second active compound can be administered simultaneously, sequentially, or independently with any of the compounds encompassed by general formulas I, II or III, and pharmaceutical salts thereof.

For the purposes of the present invention, "active compound or active ingredient" should be taken synonymously and means a chemical entity that exerts a therapeutic effect upon administration to humans or animals. Said second active compound is an insulin sensitizer, a bile acid regulator, an inhibitor of de novo lipogenesis, a hypolipidemic agent, an antioxidant, an anti-inflammatory agent, an immunomodulatory component, an anti-apoptotic agent, a gut microbiome modulating agent. agents, or antifibrotic agents.

For purposes of this invention, insulin sensitizers include, but are not limited to, PPAR agonists, incretin analogues (GLP-1 receptor agonists), DPP-4 inhibitors, SGLT2 inhibitors, ACE inhibitors, or angiotensin Includes II receptor antagonists (antihypertensives). Bile acid modulating agents include but are not limited to farnesoid X receptor agonists. Inhibitors of de novo adipogenesis include, but are not limited to, stearoyl-CoA desaturase or acetyl-CoA carboxylase inhibitors. Lipid-lowering agents include but are not limited to statins, fibrates or lipase inhibitors. Antioxidants include but are not limited to vitamin E or cysteamine. Anti-inflammatory agents include, but are not limited to, TNF-α inhibitors. Immunomodulatory components include, but are not limited to, IκB inhibitors, inflammatory chemokines antagonists (CCR2/CCR5 inhibitors) or VAP1 inhibitors. Anti-apoptotic agents include but are not limited to caspase inhibitors or ASK1 inhibitors. Gut microbiome modulating agents include, but are not limited to, antibiotics, anti-LPS IgG-rich extracts, or faecal microbial transplants. Antifibrotic agents include, but are not limited to, galectin-3 inhibitors or LOXL2 blockers.

The present invention is also for use in the alleviation and/or prevention of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), and/or symptoms and/or medical conditions associated therewith. of at least one compound of formula I, or a food acceptable salt thereof, and at least one food acceptable excipient:

where independently
R ₁ is selected from linear or cyclic mono- or dialkylamines; OR ₉ , aminoalkyl alcohols, or aminoalkyl ethers;
R ₂ is selected from benzene or heterocycle;
R ₃ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; or a benzyl group;
R ₄ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; a hydroxy or alkoxy radical; or halogen;
and _R9 are alkyl groups.

Additionally disclosed is the use of a dietary supplement as described above comprising an effective amount of at least one compound of Formula I and at least one pharmaceutically acceptable excipient, wherein:
—R ₁ is NH—(CH ₂ ) _n —NH ₂ , NH—(CH ₂ ) _n —N(CH ₃ ) ₂ (where n is a value of 0 to 4); HNCH ₂ CH ₂ OH; HNCH ₂ CH ₂ OCH ₃ ; NH—N═CH-phenyl-R ₇ ; or a cyclic amine selected from:

- _R2 is selected from

-R ₃ is selected from H, methyl, or benzyl;
-R ₄ is selected from H, methyl, methoxy, or fluorine;
-R ₇ is H or p-NO ₂ ;
—R ₈ is H, OH, or methoxy; and
—R ₅ is H, OH, or methoxy.

The present invention, as previously disclosed, is a compound of general formula I, II or III, and pharmaceutical salts thereof, or pharmaceutical compositions containing them.

The present invention further includes a method of preventing and/or treating a subject suffering from, or suffering from symptoms associated with and/or medical conditions associated with, NAFLD or NASH, the method comprising an effective amount of general formula I, administering to said subject any of the compounds encompassed by II or III, and pharmaceutically or food acceptable salts thereof, or a pharmaceutical composition, food additive or dietary supplement containing them include.

The present invention further includes a method of preventing and/or treating a subject suffering from, or suffering from symptoms associated with and/or medical conditions associated with, NAFLD or NASH, the method comprising: administering to said subject a pharmaceutical composition comprising any of the encompassed compounds, and pharmaceutically acceptable salts thereof.

Additionally, the present invention further includes a method of preventing and/or treating a subject suffering from or suffering from symptoms associated with NAFLD or NASH and/or conditions associated therewith, the method comprising: administering to said subject a functional food additive or dietary supplement comprising any of the compounds encompassed by II or III and a food acceptable salt thereof.

The most preferred compounds for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or pathologies associated therewith, according to the present invention are selected from the following compounds: , 4a, 5a, 7a, 17a, 17b, 17c, 21a, 21b, 21c, 21d, 21e, 21f, 23a, 23b, 23c, 23d, 23e, 23f, 26a, or 26b.

The present invention further provides 4a, 5a, 7a, 17a, 17b, 17c shown in Table 1 for use in the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or pathologies associated therewith. , 21a, 21b, 21c, 21d, 21e, 21f, 23a, 23b, 23c, 23d, 23e, 23f, 26a, or 26b, or a pharmaceutical composition comprising the same.

More preferred are compounds independently selected from 4a, 5a and 7a, or A pharmaceutical composition comprising

More preferred are 4a, 26a, 21a, 26b, 21b, 21c, 21e, 21d, 17a for use in the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or pathologies associated therewith. , 17c, 17b, or 21f, or a pharmaceutical composition comprising the same.

More preferred are 4a, 5a, 7a, 23b, 23c, 26a, 23a, 23e, 23d for use in the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or pathologies associated therewith. , 26b, 21d, 17a, 17c, 17b, or 23f, or a pharmaceutical composition comprising the same.

More preferred is independently selected from 4a, 26a, 17a, 17c, or 17b for use in the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or pathologies associated therewith compound, or a pharmaceutical composition comprising the same.

More preferred is a compound independently selected from 4a or 5a, or a medicament containing it, for use in the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or pathologies associated therewith composition.

More preferably, the compound for use according to the invention is the hydrochloride salt of 4a or 5a. More preferably, a compound independently selected from 4a, or a pharmaceutical composition comprising it, for use in the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or pathologies associated therewith be. More preferably, the compound for use according to the invention is the hydrochloride salt of 4a.

Additionally, the present invention includes methods of preventing and/or treating a subject suffering from or suffering from symptoms associated with NAFLD or NASH and/or suffering from associated medical conditions, which methods are presented in Table 1. an effective amount of 4a, 5a, 7a, 17a, 17b, 17c, 21a, 21b, 21c, 21d, 21e, 21f, 23a, 23b, 23c, 23d, 23e, 23f, 26a, or 26b, or administering to said subject a pharmaceutical composition, food additive, or dietary supplement comprising More preferably, an effective amount of 4a, 5a, and 7a; or 4a, 26a, 21a, 26b, 21b, 21c, 21e, 21d, 17a, 17c, 17b, or 21f; 23c, 26a, 23a, 23e, 23d, 26b, 21d, 17a, 17c, 17b, or 23f; or 4a, 26a, 26b, 17a, 17c, or 17b; or 4a or 5a to the subject. .

More preferably, the invention includes a method of preventing and/or treating a subject suffering from, or suffering from symptoms associated with, and/or medical conditions associated with, NAFLD or NASH, said method comprising an effective amount of 4a , or a pharmaceutical composition, food additive, or dietary supplement comprising the same to said subject.

The present invention further comprises optionally any inactive ingredients, carriers, excipients, etc. for use in the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or medical conditions associated therewith. , independently of 4a, 5a, 7a, 17a, 17b, 17c, 21a, 21b, 21c, 21d, 21e, 21f, 23a, 23b, 23c, 23d, 23e, 23f, 26a or 26b as shown in Table 1 or pharmaceutically or food acceptable salts thereof and combinations thereof, including pharmaceutical compositions, functional food additives, or dietary supplements. more preferably from 4a, 5a and 7a; or from 4a, 26a, 21a, 26b, 21b, 21c, 21e, 21d, 17a, 17c, 17b or 21f; at least one independently selected from 26a, 23a, 23e, 23d, 26b, 21d, 17a, 17c, 17b, or 23f; or from 4a, 26a, 26b, 17a, 17c, or 17b; or from 4a or 5a containing one compound.

In preferred embodiments, the present invention provides any inert ingredient, carrier, or excipient for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or pathologies associated therewith. pharmaceutical compositions, functional food additives, or dietary supplements containing 4a, and pharmaceutically or food acceptable or acceptable salts thereof and combinations thereof, with agents, etc.

The present invention also provides all of the compounds listed above in Table 1 in the synthesis of compounds for use in the prevention and/or treatment of NAFLD or NASH and symptoms associated therewith and/or pathologies associated therewith. Includes intermediate compounds.

In particular, the invention provides 1a, 1b, 2a, 2b, 3a, 3b, 6a, 6b, 7b, 8, 13a, 13b, 13c, 14a, 14b, 14c, 15a, 15b, 15c, 16a, 16b, 16c, 18a, 18b, 18c, 18d, 18e, 18f, 19a, 19b, 19c, 19d, 19e, 19f, 20a, 20b, 20c, 20d, 20e, 20f, 22a, 22b, 22c, 22d, 22e, 22f, 24a, 24b, 25a, or 25b, and the invention further includes NAFLD or A method for preventing and/or treating a subject suffering from NASH, or suffering from symptoms and/or medical conditions associated therewith, said method comprising said intermediate compound, or a pharmaceutical composition, food comprising the same administering an additive, or dietary supplement, to said subject.

In particular, the present invention encompasses intermediate compounds selected from: 7b, 15a, 15b for use in the prevention and/or treatment of NAFLD or NASH, and symptoms associated therewith and/or pathologies associated therewith. , 15c, 16a, 16b, 16c, 19a, 19b, 19c, 19d, 19e, 19f, 20a, 20b, 20c, 20d, 20e, 20f, 22a, 22b, 22c, 22d, 22e, 22f, 24a, 24b, 25a , or 25b. and the invention further includes a method for preventing and/or treating a subject suffering from or suffering from symptoms associated with NAFLD or NASH and/or suffering from a medical condition associated with said intermediate compound, or administering to said subject a pharmaceutical composition, food additive, or dietary supplement comprising the same.

In particular, the present invention provides 7b, 16a, 16b, 16c, 20a, 20b, 20c, 20d; 20e, 20f, 22a, 22b, 22c, 22d, 22e, 22f, 24a, 24b, 25a, or 25b, and the invention further encompasses intermediate compounds suffering from or associated with NAFLD or NASH. A method for preventing and/or treating a subject suffering from a symptom of and/or an associated medical condition, the method comprising adding the intermediate compound, or a pharmaceutical composition, food additive, or dietary supplement containing administering to said subject.

Preferably, the present invention also provides compounds named 4a, 5a, 7a, 21a, 21b, 21e, 23a, 23b, 23d, 23e, 23f, or 26b, alone or in combination, or pharmaceutical compositions comprising the same for use in the treatment or prevention of NAFLD or NASH and symptoms and/or pathologies associated therewith, or for the treatment of NAFLD or NASH and symptoms and/or pathologies associated therewith It is particularly suitable for use in the manufacture of a medicament for

The invention further relates to the above-disclosed general formula I, for use as a functional food additive or dietary supplement, particularly for preventing or reducing symptoms associated with NAFLD or NASH, and medical conditions associated therewith. Any of the compounds encompassed by II or III, or functional food additives or dietary supplements containing same.

Preferably, the present invention also relates to the compounds named 4a, 5a, 7a, 21a, 21b, 21e, 23a, 23b, 23d, 23e, 23f, or 26b, or functional foods comprising the same, either alone or in combination. additives or dietary supplements, which are particularly suitable for use as functional food additives or dietary supplements to specifically prevent or reduce symptoms associated with NAFLD or NASH, and medical conditions associated therewith. ing.

Additionally, the present invention also includes methods for preventing or reducing symptoms associated with NAFLD or NASH, and pathologies associated therewith, in patients suffering from said symptoms and associated pathologies, said methods comprising: administering to said subject an effective amount of any of the compounds encompassed by general formulas I, II or III disclosed in , or a functional food additive or dietary supplement comprising the same.

Compounds 4a (SJT4A) and 5a (SJT5A), all at doses of 50 mg/kg, to controls treated with saline solution (0.9% NaCl) and positive controls treated with metformin (150 mg/kg), and In vivo effect of the hydrochloride salt of compound 7a (SJT7A) on reducing insulin resistance (HOMA-IR) in DIO model mice during 36 days of treatment. In vivo effects of compounds of the invention in reducing liver fat. Hydrochloride salt of compound 4a (SJT4A) and compound 7a (SJT7A) for 36 days compared to controls treated with saline solution (0.9% NaCl) and positive controls treated with metformin (150 mg/kg) Cholesterol liver content in DIO mice treated with (both 50 mg/kg). In vivo effects of compounds of the invention in reducing liver fat. Hydrochloride salt of compound 4a (SJT4A) and compound 7a (SJT7A) for 36 days compared to controls treated with saline solution (0.9% NaCl) and positive controls treated with metformin (150 mg/kg) Shown are hepatic triglyceride levels in DIO mice treated with (both 50 mg/kg). In vivo effects of compounds of the invention in reducing liver fat. Hydrochloride salt of compound 4a (SJT4A) and compound 7a (SJT7A) for 36 days compared to controls treated with saline solution (0.9% NaCl) and positive controls treated with metformin (150 mg/kg) Liver fatty acid levels in DIO mice treated with (both 50 mg/kg). Figure 4 shows the in vivo effect of compound 4a (SJT4a) hydrochloride on liver lipid content. DIO-NASH mice (C57B1/6J male mice) were compared with diseased controls of DIO-NASH mice and non-diseased control LEAN-CHOW mice co-treated with vehicle (0.9% NaCl). Treated with compound 4a hydrochloride (50 mg/kg, po, twice daily) for 8 weeks. Representative images of liver morphology for the three mice at the end of treatment are shown (magnification x20). Figure 3 shows the in vivo effect of the hydrochloride salt of compound 4a (SJT4a) on hepatic triglyceride content. Figure 3 shows the in vivo effect of the hydrochloride salt of compound 4a (SJT4a) on hepatic triglyceride content. In both cases, DIO-NASH mice (C57B1/6J male mice) were compared with diseased controls and non-diseased controls LEAN-NASH mice, both treated with vehicle (0.9% NaCl). Compared to CHOW mice, treated with the hydrochloride salt of compound 4a (50 mg/kg, po, twice daily) for 8 weeks. Compound 4a at both 50 mg/kg in DIO mice after 36 days of treatment compared to controls treated with saline solution (0.9% NaCl) and positive controls treated with metformin (150 mg/kg) In vivo effects of hydrochloride salts of (SJT4A) and 5a (SJT5A) to reduce liver overweight associated with fatty liver. Figure 4 shows the in vivo effect of the hydrochloride salt of compound 4a (SJT4a) on total liver weight. DIO-NASH mice (C57B1/6J male mice) versus diseased controls and non-diseased control LEAN-CHOW mice, both treated with vehicle (0.9% NaCl). , with the hydrochloride salt of Compound 4a (50 mg/kg, po, twice daily) for 8 weeks. Figure 4 shows the in vivo effects of the hydrochloride salt of compound 4a (SJT4a) in absolute body weight. DIO-NASH mice (C57B1/6J male mice) versus diseased controls and non-diseased control LEAN-CHOW mice, both treated with vehicle (0.9% NaCl). , with the hydrochloride salt of Compound 4a (50 mg/kg, po, twice daily) for 8 weeks. Figure 4 shows the in vivo effects of the hydrochloride salt of Compound 4a (SJT4a) on relative body weights. DIO-NASH mice (C57B1/6J male mice) versus diseased controls and non-diseased control LEAN-CHOW mice, both treated with vehicle (0.9% NaCl). , with the hydrochloride salt of Compound 4a (50 mg/kg, po, twice daily) for 8 weeks. Figure 3 shows the in vivo effect of compound 4a (SJT4a) hydrochloride on plasma ALT (alanine aminotransferase). DIO-NASH mice (C57B1/6J male mice) were compared against diseased controls and non-diseased controls LEAN-CHOW of DIO-NASH mice both treated with vehicle (0.9% NaCl). Treated with the hydrochloride salt of compound 4a (50 mg/kg, po, twice daily) for 8 weeks. Figure 3 shows the in vivo effect of compound 4a (SJT4a) hydrochloride on plasma AST (aspartate aminotransferase). DIO-NASH mice (C57B1/6J male mice) were compared against diseased controls and non-diseased controls LEAN-CHOW of DIO-NASH mice both treated with vehicle (0.9% NaCl). Treated with the hydrochloride salt of compound 4a (50 mg/kg, po, twice daily) for 8 weeks. 50 mg/kg orally twice daily for 8 weeks (A) LEAN-CHOW mice treated with vehicle, (B) DIO-NASH mice treated with vehicle, or (C) Compound 4a (SJT4a) In vivo assay to measure the NAFLD activity score (NAS) in DIO-NASH mice treated with the hydrochloride of The in vivo effect of the hydrochloride salt of compound 4a (SJT4a) on liver fibrosis was shown and collagen type I (col1a1) was measured as a fibrosis marker. Liver samples were administered for 8 weeks (50 mg/kg po dosing, twice daily), from DIO-NASH mice (C57Bl/6J male mice) treated with the hydrochloride salt of compound 4a. Representative images of liver samples for each of the mouse groups stained with anti-type I collagen at the end of the study are shown (magnification 20x). The in vivo effect of compound 4a (SJT4a) hydrochloride on hepatitis was shown and galectin-3 was measured as a hepatitis marker. Liver samples were administered for 8 weeks (50 mg/kg po dosing, twice daily), from DIO-NASH mice (C57Bl/6J male mice) treated with the hydrochloride salt of compound 4a. Representative images of liver samples for each of the groups of mice stained with anti-galectin-3 at the end of the study are shown (magnification 20x). Compound 4a (SJT4a) versus diseased control and non-diseased control LEAN-CHOW mice in DIO-NASH mice both treated with vehicle (0.9% NaCl) in collagen gene expression as a fibrosis marker. Effect of 8 weeks treatment of DIO-NASH mice with 50 mg/kg hydrochloride of: (A) type I collagen-alpha 1 (COL1A1) and (B) alpha 2 (COL1A2) and (C) type III collagen alpha 1 ( COL3A1). Compound 4a (SJT4a) versus diseased control and non-diseased control LEAN-CHOW mice in DIO-NASH mice both treated with vehicle (0.9% NaCl) in collagen gene expression as a fibrosis marker. (A) collagen V alpha 1 (COL5A1), (B) alpha 2 (COL5A2) and (C) alpha 3 (COL5A3), (D ) Type IV collagen alpha 1 (COL6A1), (E) alpha 2 (COL6A2) and (F) alpha 3 (COL6A3).

Example 1: In vivo efficacy of compounds of the invention in reducing insulin resistance.

As described herein above, insulin resistance is recognized as an important development or symptom associated with the development of NAFLD.

To evaluate the performance of compounds of the present disclosure in reducing insulin resistance, compared to controls treated with saline solution (0.9% NaCl) and DIO mice treated with metformin as a positive control, DIO mice (C57B1/6J male mice) were treated with compounds 4a, 5a hydrochloride; and compound 7a for 36 days.

For evaluation, 60 C57B1/6J male mice from Charles River France (16 weeks old at time of shipment, average weight of 40 g) were fed a high fat diet for 10 weeks. Animals were housed in ventilated and enriched cages (310×125×127 mm ³ ) throughout the course of the experiment. The animal's cage bedding was changed at least once a week. Mice were housed in groups of 10 mice at 22±2° C. and 50±10% relative humidity under a normal 12 hour light cycle (lights switched off at 08:00 pm). After acceptance, the mice were kept for an additional 3 weeks for acclimatization. High fat diet (D12492 purchased from Research Diet Inc.; 60% fat) and tap water were provided ad libitum during the entire study (acclimatization + treatment phase).

After an acclimation period, mice were fasted for 6 hours and blood glucose and plasma insulin were measured. Ten animals with lower blood glucose/plasma insulin were discarded from the study. The other was then randomized into 5 homogeneous groups (n=10 mice per group) according to blood glucose/plasma insulin levels (HOMA-IR or Homeostasis Model Assessment-Insulin Resistance Index) and body weight. Mice were treated orally twice daily with saline solution (control group), test compound (4a, 5a, 7a; 50 mg/kg) or metformin (150 mg/kg) for 36 days.

Body weight was measured weekly along with blood glucose and plasma insulin after a 6 hour fast.

Blood glucose was measured by sampling blood from the tip of the tail: a drop of blood is taken and placed on a glucometer plate (Accu-Check R glucometer, Roche, Switzerland). Plasma insulin was detected by ELISA ((Elisa Kit, Eurobio, France) using 5 μl samples.

The Homeostasis Model Assessment-Insulin Resistance Index (HOMA-IR), used to quantify insulin resistance, was calculated from fasting blood glucose and plasma insulin levels were calculated as follows: HOMA-IR = (mM glucose X μU/mL insulin)/22.5, taking this constant as the normalization factor (Matthews et al., 1985, Diabetologia 28(7), 412-9).

FIG. 1 shows HOMAR-IR results for mice treated with each of the compounds tested, mice treated with metformin, and control animals after 1, 8, 15, 22, 29, and 36 days. Mice treated with the hydrochloride salts of compounds 4a and 5a, and compound 7a exhibited insulin resistance that was lower compared to control animals and similar to metformin-treated animals over the entire period of treatment.

Data are mean±sem from 10 animals. e. m. shown as a value.

Example 2: In vivo effects of compounds of the invention on hepatic lipid content associated with fatty liver.

2.1. Effect of compounds 4a and 7a on hepatic lipid content: 36-day assay in DIO mice.
To evaluate the performance of compounds of the present disclosure in reducing hepatic lipid content, DIO mice treated with saline solution (0.9% NaCl) as a control, and DIO mice treated with metformin as a positive control, DIO mice (C57B1/6J male mice) were treated with compound 4a hydrochloride; and compound 7a for 36 days. Animals were housed in ventilated and enriched cages (310×125×127 mm ³ ) throughout the course of the experiment. The animal's cage bedding was changed at least once a week. Mice were housed in groups of 10 mice at 22±2° C. and 50±10% relative humidity under a normal 12 hour light cycle (lights switched off at 08:00 pm). After acceptance, the mice were kept for an additional 3 weeks for acclimatization. High fat diet (D12492 purchased from Research Diet Inc.; 60% fat) and tap water were provided ad libitum during all assessments (acclimatization + treatment phase).

After an acclimation period, mice were fasted for 6 hours and blood glucose and plasma insulin were measured. Ten animals with lower blood glucose/plasma insulin were discarded from the study. The other was then randomized into 5 homogeneous groups (n=10 mice per group) according to blood glucose/plasma insulin levels (HOMA-IR) and body weight. Mice were treated orally twice daily with saline solution (control group), SJT test compounds (4a, 7a; 50 mg/kg) or metformin (150 mg/kg) for 36 days.

Finally, mice were sacrificed and livers were harvested, weighted and stored at -80°C for further analysis. Liver samples were dissected for liver lipid assay. Miao et al. , J Lipid Res, 2004, 45, 1410-17, liver lipids were analyzed from homogenates of liver samples using a colorimetric commercial assay kit after lipid solubilization in deoxycholate. analyzed. The sample was homogenized with an ultrasonic probe with 500 μl of distilled water for just a few seconds. Cholic acid was then added at 1% to solubilize the lipids. Various lipid measurements were then performed using a colorimetric kit from Sobioda, France.

Figures 2A, 2B, and 2C show the levels of liver cholesterol, liver triglycerides, and liver fatty acids, respectively, after 36 days of treatment with compound 4a hydrochloride and compound 7a. In each case, both groups of mice treated with compounds 4a and 7a showed lower lipid levels than untreated control animals. Data are mean±sem from 10 animals. e. m. shown as a value.

2.2. Effect of compound 4a hydrochloride on hepatic lipid content: an 8-week assay in DIO-NASH mice.
The DIO-NASH mouse model is fed a high-fat diet that results in non-alcoholic fatty liver disease, and C57B1/6J mice are fed a 40% high-fat diet pre-assay. For evaluation of the effects of compounds of the invention on hepatic lipid content in non-alcoholic fatty liver disease, 34 C57B1/6J male mice (5 weeks old at time of shipment) from Janvier, France were used. A group of DIO-NASH mice were fed a high-fat diet (40% AMLN diet, D09100301 Research Diets, USA) and a control group (LEAN-CHOW) were fed a normal solid diet (Altromin 1324, Brogaarden, USA) for 35 weeks prior to testing. Denmark).

Animals (housed in single housing) were checked at least once daily for signs of abnormal behavior, abnormal locomotor activity, ataxia, or clinical signs of disease (lack of grooming, puffy fur, handling). Occasional signs of pain, excessive weight loss) were followed closely. Health status that warranted additional evaluation was examined by a clinical veterinarian or by a professional working under the supervision of a clinical veterinarian. The same abnormal behavior and clinical symptoms of disease were used to determine if an animal was not growing during the study period and should be terminated for ethical reasons. Possible treatments recommended by the veterinarian were performed after agreement with the study director. Mice were housed on a regular 12 hour light cycle (lights off at 3 pm) and the room environment was controlled (target range: temperature 21±2° C.; relative humidity 50±1%).

Each animal was uniquely identified by an implantable microchip (Pet ID Microchip, E-vet) upon arrival at the animal unit. Animals were identified using a WS-1 weigh station (MBrose, Denmark) connected to a laptop running HM02Lab software (Ellegaard Systems, Denmark). The HM02Lab software matches body weights with animal IDs. For liver biopsies, mice were anesthetized by inhalation anesthesia using isoflurane (2-3%). A small abdominal incision was made in the midline to expose the left lateral lobe of the liver. A cone-shaped wedge (approximately 50 mg) of liver tissue was excised from the distal portion of the lobe and fixed in 10% neutral buffered formalin (4% formaldehyde) for histology. Sections of the liver were immediately electrocoagulated using a bipolar coagulator (ERBE VIO 100 electrocautery). The liver is returned to the abdominal cavity, the abdominal wall is sutured, and the skin is closed with staples. For post-operative recovery, mice received carprofen (5 mg/kg) administered subcutaneously on the day of surgery and on days 1 and 2 post-surgery. Test samples were stored at -80°C prior to histological examination.

2.2. A. Total Fat Content in Liver To assess the effect of compound 4a on liver lipid content in mice with non-alcoholic fatty liver disease, we measured the total fat content of DIO-NASH mice treated with saline solution (0.9% NaCl). DIO-NASH mice (C57B1/6J male mice) were treated with compound 4a hydrochloride for 8 weeks versus control and LEAN-CHOW mice that were additionally treated with saline solution (0.9% NaCl) as a positive control. (50 mg/kg po, twice daily). Liver lipid content was determined by morphometry. As previously mentioned, a group of DIO-NASH mice were fed a high-fat diet (40% AMLN diet, D09100301 Research Diets, USA) and a control group (LEAN-CHOW) was fed a normal chow diet for 35 weeks prior to testing. Chow (Altromin 1324, Brogaarden, Denmark) was provided.

FIG. 3A shows that at the end of the assay, liver content increased in vehicle-treated DIO-NASH mice control group (middle column) compared to vehicle-treated LEAN-CHOW mice control group (left column). The group of DIO-NASH mice treated with the hydrochloride salt of compound 4a (right column) showed decreased liver lipid content (reduced steatosis) compared to the control group of DIO-NASH mice. rice field.

Data are mean±sem from 10-12 animals. e. m. shown as a value. ***p<0.001 vs. DIO-NASH vehicle; one-way ANOVA with Dunnett's multiple comparison test.

Figure 3B shows representative images of liver morphology for the three groups of mice at the end of treatment (magnification x20), where compound 4a was compared to the vehicle-treated DIO-NASH mouse control group. It can be seen that treatment with DIO reduced steatosis in DIO-NASH mice. Liver samples were stained with Hematoxylin & Eosin (H&E). To that end, samples were incubated with Mayer's Hematoxylin (Dako), washed with tap water, stained with Eosin Y solution (Sigma-Aldrich), hydrated, mounted in Pertex, and then dried prior to scanning. .

2.2. B. Liver Content of Triglycerides and Cholesterol To assess the effect of compound 4a on triglycerides or cholesterol in animals with non-alcoholic fatty liver disease, DIO-NASH mice treated with both vehicle (0.9% NaCl) DIO-NASH mice (C57Bl/6J male mice) were treated for 8 weeks with the hydrochloride salt of compound 4a (50 mg/kg, p.o. , twice a day). A group of DIO-NASH mice were fed a high-fat diet (40% AMLN diet, D09100301 Research Diets, USA) and a control group (LEAN-CHOW) were fed a normal solid diet (Altromin 1324, Brogaarden, USA) for 35 weeks prior to testing. Denmark).

Liver triglyceride content was determined on a Cobas™ C-501 automatic analyzer using triglyceride reagent (Cat. no. 22-045-795, Roche Diagnostics, Germany). Homogenized liver tissue was heated twice at 80-100° C., centrifuged in a microcentrifuge, and triglyceride content was measured in the supernatant.

FIG. 4A shows that vehicle-treated DIO-NASH mice control group (middle column) compared to vehicle-treated LEAN-CHOW mice control group (left column) in triglyceride levels at the end of the assay. shows an increase, indicating that the group of DIO-NASH mice treated with the hydrochloride salt of compound 4a (right column) showed decreased triglyceride levels compared to the control group of DIO-NASH mice.

Data are mean±sem from 10-12 animals. e. m. shown as a value. ***p<0.001 vs. DIO-NASH vehicle; one-way ANOVA with Dunnett's multiple comparison test (all columns versus DIO-NASH vehicle).

FIG. 4B shows that at the end of the assay, vehicle-treated DIO-NASH mice control group (middle column) compared to vehicle-treated LEAN-CHOW mice control group (left column) increased cholesterol levels. shows an increase, indicating that the group of DIO-NASH mice treated with the hydrochloride salt of compound 4a (right column) showed decreased cholesterol levels compared to the control group of DIO-NASH mice.

Data are mean±sem from 10-12 animals. e. m. shown as a value. **p<0.01, ***p<0.001 vs. DIO-NASH vehicle; one-way ANOVA with Dunnett's multiple comparison test (all columns versus DIO-NASH vehicle).

Example 3: In vivo effect of compounds of the invention on liver weight.

3.1: Effect of compounds 4a and 5a on liver weight: 36-day assay in DIO mice.
To assess the ability of compounds of the present disclosure to reduce liver weight, DIO mice treated with vehicle (0.9% NaCl) were compared to control DIO mice treated with metformin as a positive control. DIO mice (C57B1/6J male mice) were treated with hydrochloride salts of compounds 4a and 5a for 36 days.

For evaluation, 60 C57B1/6J male mice from Charles River France (16 weeks old at time of shipment, average weight of 40 g) were fed a high fat diet for 10 weeks. Animals were housed in ventilated and enriched cages (310×125×127 mm ³ ) throughout the course of the experiment. The animal's cage bedding was changed at least once a week. Mice were housed in groups of 10 mice at 22±2° C. and 50±10% relative humidity under a normal 12 hour light cycle (lights switched off at 08:00 pm). After acceptance, the mice were kept for an additional 3 weeks for acclimatization. High fat diet (D12492 purchased from Research Diet Inc.; 60% fat) and tap water were provided ad libitum during the entire study (acclimatization + treatment phase).

After an acclimation period, mice were fasted for 6 hours and blood glucose and plasma insulin were measured. Ten animals with lower blood glucose/plasma insulin were discarded from the study. The other was then randomized into 5 homogeneous groups (n=10 mice per group) according to blood glucose/plasma insulin levels (HOMA-IR) and body weight. Mice were treated orally twice daily for 36 days with either saline solution (control group), SJT test compounds (hydrochloride salts of compounds 4a and 5a; 50 mg/kg) or metformin (150 mg/kg).

Finally, mice were sacrificed and livers were harvested, weighted and stored at -80°C for further analysis.

FIG. 5 shows liver weight after 36 days of treatment with the hydrochloride salts of compounds 4a and 5a. Both groups of mice treated with hydrochloride salts of compounds 4a and 5a show lower liver weights than either metformin-treated mice or control animals.

Data are mean±sem from 10 animals. e. m. shown as a value.

3.2. Effect of compound 4a on liver weight: 8-week assay in DIO-NASH mice.
Animals were housed and examined under the same conditions used in Example 2.2. A liver biopsy was also performed under the same conditions as in Example 2.2.

To assess the effect of compound 4a on liver weight in mice with non-alcoholic fatty liver disease, DIO-NASH mice both treated with vehicle (0.9% NaCl) were treated with diseased controls and non-drugs. DIO-NASH mice (C57B1/6J male mice) were treated with the hydrochloride salt of compound 4a (50 mg/kg po, twice daily) for 8 weeks relative to diseased control LEAN-CHOW mice. A group of DIO-NASH mice were fed a high-fat diet (40% AMLN diet, D09100301 Research Diets, USA) and a control group (LEAN-CHOW) were fed a normal solid diet (Altromin 1324, Brogaarden, USA) for 35 weeks prior to testing. Denmark).

FIG. 6 shows total liver weights at the end of the assay, where the vehicle-treated DIO-NASH mouse control group (middle column) and the vehicle-treated LEAN-CHOW mouse control group (left column). column), and the group of DIO-NASH mice treated with the hydrochloride salt of compound 4a (right column) showed a decrease in liver weight compared to the control group of DIO-NASH mice. .

Data are mean±sem from 10-12 animals. e. m. shown as a value. ***p<0.001 vs. DIO-NASH vehicle; one-way ANOVA with Dunnett's multiple comparison test (all columns versus DIO-NASH vehicle).

Example 4: In vivo effects of compound 4a on absolute and relative body weight.

An assay was performed to evaluate the effects of compounds of the present invention on body weight in animals with non-alcoholic fatty liver disease (DIO-NASH mice). For this assay, the effects of compound 4a on absolute and relative body weight were evaluated. DIO-NASH mice (C57B1/6J male mice) versus diseased controls and non-diseased control LEAN-CHOW mice, both treated with vehicle (0.9% NaCl). , with the hydrochloride salt of Compound 4a (50 mg/kg, po, twice daily) for 8 weeks. A group of DIO-NASH mice were fed a high-fat diet (40% AMLN diet, D09100301 Research Diets, USA) and a control group (LEAN-CHOW) were fed a normal solid diet (Altromin 1324, Brogaarden, USA) for 35 weeks prior to testing. Denmark).

Body weights were taken daily for each of the groups. Animals were maintained under the same conditions as used in Example 2.2.

Figure 7A shows the absolute body weights measured for each of the three animal groups. Vehicle-treated DIO-NASH mice demonstrated increased body weight compared to vehicle-treated LEAN-CHOW mice, and treatment with the hydrochloride salt of compound 4a significantly reduced vehicle-treated DIO-NASH mice. body weight loss in DIO-NASH mice when compared to DIO-NASH mice.

Data are mean±sem from 10-12 animals. e. m. shown as a value. **P<0.01, ***P<0.001 vs. NASH vehicle. One-way ANOVA with Dunnett's multiple comparison test (versus NASH vehicle) performed on day 54 of treatment.

FIG. 7B shows the percentage of relative body weight measured for each of the three animal groups relative to body weight at the start of treatment (100% on day 0). Although vehicle-treated DIO-NASH and LEAN-CHOW animals did not show significant body weight changes during vehicle treatment, treatment of DIO-NASH mice with compound 4a hydrochloride increased the relative body weight was significantly reduced.

Data are mean±sem from 10-12 animals. e. m. shown as a value. ***P<0.001 vs. NASH vehicle. One-way ANOVA with Dunnett's multiple comparison test (versus NASH vehicle) performed on day 54 of treatment.

Example 5: In vivo effect of compound 4a on liver toxicity

An assay was performed to evaluate the effects of compounds of the invention on liver toxicity in animals with non-alcoholic fatty liver disease (DIO-NASH mice). This assay evaluated the effects of compound 4a on plasma alanine aminotransferase (ALT) and plasma aspartate aminotransferase (AST). DIO-NASH mice (C57B1/6J male mice) versus diseased controls and non-diseased control LEAN-CHOW mice, both treated with vehicle (0.9% NaCl). , with the hydrochloride salt of Compound 4a (50 mg/kg, po, twice daily) for 8 weeks. Animals were maintained under the same conditions as used in Example 2.2. A group of DIO-NASH mice were fed a high-fat diet (40% AMLN diet, D09100301 Research Diets, USA) and a control group (LEAN-CHOW) were fed a normal solid diet (Altromin 1324, Brogaarden, USA) for 35 weeks prior to testing. Denmark).

To assess alanine transaminase (ALT) and aspartate transaminase (AST), blood samples were collected into heparinized tubes, plasma separated and stored at -80°C until analysis. ALT and AST were measured using commercially available kits (Roche Diagnostics, Germany) on a Cobas™ C-501 automatic analyzer according to the manufacturer's instructions.

FIG. 8A shows plasma ALT (U/L) for each of the three animal groups at the end of treatment. The DIO-NASH control group treated with vehicle showed an increase in plasma ALT compared to LEAN-CHOW animals. Treatment with the hydrochloride salt of compound 4a lowered plasma ALT levels in DIO-NASH animals when compared to vehicle-treated DIO-NASH animals.

Data are mean±sem from 10-12 animals. e. m. shown as a value. ***P<0.001 vs. NASH vehicle. One-way ANOVA with Dunnett's multiple comparison test (all columns versus NASH vehicle).

FIG. 8B shows plasma AST (U/L) for each of the three animal groups at the end of treatment. The DIO-NASH control group treated with vehicle showed an increase in plasma AST compared to LEAN-CHOW animals. Treatment with the hydrochloride salt of compound 4a lowered plasma AST levels in DIO-NASH animals when compared to vehicle-treated DIO-NASH animals.

Data are mean±sem from 10-12 animals. e. m. shown as a value. ***P<0.001 vs. NASH vehicle. One-way ANOVA with Dunnett's multiple comparison test (all columns versus NASH vehicle).

Example 6: In vivo effect of compound 4a on liver fibrosis and hepatitis as measured by NAFLD activity score (NAS)

Non-alcoholic fatty liver disease (NAFLD) activity score or NAS was assessed by measuring liver steatosis, lobular inflammation, balloon degeneration and fibrosis. The total NAS score represents the sum of scores for adiposity, inflammation, and ballooning and ranges from 0-8 as follows:

The % adiposity refers to the amount of surface area of the sample covered by adiposity as assessed by the low to medium power test.

Values given to measure inflammation correspond to the number of inflammatory foci per field using 200x magnification. Foci are defined as clusters, not rows, of >3 inflammatory cells. Eosinophilic cell bodies are not included in the assessment of inflammation.

The balloon degeneration value is measured by the amount of balloon cells corresponding to degenerated hepatocytes with cytoplasm removed, swollen, swollen, rounded and reticulated cytoplasm.

To assess the effect of compound 4a on NAS in mice with non-alcoholic fatty liver disease, diseased controls and non-diseased DIO-NASH mice, both treated with vehicle (0.9% NaCl), were analyzed. DIO-NASH mice (C57B1/6J male mice) were treated with the hydrochloride salt of compound 4a (50 mg/kg, po, twice daily) for 8 weeks relative to normal control LEAN-CHOW mice. Animals were housed and examined under the same conditions used in Example 2.2. A liver biopsy was also performed under the same conditions as in Example 2.2. A group of DIO-NASH mice were fed a high-fat diet (40% AMLN diet, D09100301 Research Diets, USA) and a control group (LEAN-CHOW) were fed a normal solid diet (Altromin 1324, Brogaarden, USA) for 35 weeks prior to testing. Denmark).

Liver samples were formalin-fixed, paraffin-embedded, and sections stained with hematoxylin-eosin (H&E) and Sirius red. Kleiner et al. Samples are scored for NAS and fibrosis using the clinical criteria outlined by 2005.

FIG. 9 shows NAS score changes in pre- and post-study liver biopsies for each of the groups of animals. For each animal, a line indicates the change from pre-study biopsy to post-study biopsy.

FIG. 9A shows no significant change in the NAS scores of the LEAN-CHOW group (vehicle-treated controls), where all animals show low scores both before and after the test. FIG. 9B shows how DIO-NASH mice exhibit higher score characteristics before testing and how scores tend to increase after testing when compared to the scores of LEAN-CHOW mice. show. FIG. 9C shows how DIO-NASH mice treated with the hydrochloride salt of compound 4a generally had decreased NAS scores after treatment.

To assess the efficacy of the compounds of the present invention on liver fibrosis in the treatment of non-alcoholic fatty liver disease, collagen type I (fibrosis) levels in the liver were measured after an 8-week study, and DIO -NASH mice (C57B1/6J male mice) versus DIO-NASH mice diseased controls and non-diseased control LEAN-CHOW mice, both treated with vehicle (0.9% NaCl) Treated with the hydrochloride salt of compound 4a (50 mg/kg, po, twice daily) for 8 weeks.

Collagen type I content was measured by staining with IHC stain: Collagen type I (Southern Biotech, Cat. 1310-01) IHC was performed using standard procedures. Briefly, after antigen retrieval and blocking of endogenous peroxidase activity, primary antibodies were used to incubate liver slides. Primary antibodies were detected using a biotinylated secondary antibody and amplified using the vectastain-TSA-vectastain method, a polymeric HRP linker antibody conjugate. The primary antibody was then visualized with DAB as a chromogen. Finally, the sections were counterstained in hematoxylin and slipped over.

FIG. 10A shows quantification of the total content of hepatic type I collagen (col1a1) determined by morphometry in three groups of mice. The vehicle-treated DIO-NASH control group of mice demonstrated an increase in liver col1a1 compared to the vehicle-treated LEAN-CHOW control group.
DIO-NASH mice treated with the hydrochloride salt of compound 4a showed a decrease in total liver col1a1 compared to the vehicle-treated DIO-NASH control group.

Data are mean±sem from 10-12 animals. e. m. shown as a value. *P<0.05, ***P<0.001 vs. NASH vehicle. One-way ANOVA with Dunnett's multiple comparison test (all columns versus NASH vehicle).

FIG. 10B shows representative images of liver samples for each of the groups of mice stained with anti-type I collagen (col1a1) at the end of the study (magnification 20×). The images show visible changes in this fibrosis marker after DIO-NASH animals were treated with the hydrochloride salt of Compound 4a, confirming the results in Figure 10A.

To assess the effects of compounds of the invention on liver inflammation in the treatment of non-alcoholic fatty liver disease, liver galactin-3 (a marker of inflammation) levels were determined after an 8-week study, wherein DIO-NASH Mice (C57Bl/6J male mice) were treated with compound 4a against diseased controls and non-diseased control LEAN-CHOW mice, both DIO-NASH mice treated with vehicle (0.9% NaCl). hydrochloride (50 mg/kg, po, twice daily) for 8 weeks.

Galectin-3 was measured by galectin-3 IHC staining: Galectin-3 (Biolegend, Cat. #125402) IHC was performed using standard procedures. Briefly, after antigen retrieval and blocking of endogenous peroxidase activity, slides were incubated with primary antibody. Primary antibodies were detected using a linker secondary antibody and subsequently amplified using a polymeric HRP linker antibody conjugate.
The primary antibody was then visualized with DAB as a chromogen. Finally, the sections were counterstained in hematoxylin and slipped over.

FIG. 11A shows quantification of total hepatic galectin-3 content as determined by morphometry in three groups of mice. The vehicle-treated DIO-NASH control group of mice demonstrated increased galectin-3 content compared to the vehicle-treated LEAN-CHOW control group. DIO-NASH mice treated with the hydrochloride salt of compound 4a showed decreased total galectin-3 content compared to the vehicle-treated DIO-NASH control group.

Data are mean±sem from 10-12 animals. e. m. shown as a value. ***P<0.001 vs. NASH vehicle. One-way ANOVA with Dunnett's multiple comparison test (all columns versus NASH vehicle).

FIG. 11B shows representative images of liver samples for each of the groups of mice stained with anti-galectin-3 at the end of the study (magnification 20×). The images show visible changes in this inflammatory marker after DIO-NASH animals were treated with the hydrochloride salt of Compound 4a, confirming the results in Figure 11A.

Example 7: Differential Expression Analysis: In Vivo Effects of Compound 4a.

The effects of compounds of the invention on the development of liver fibrosis were further tested by differential gene expression analysis by RNAseq, in which DIO-NASH mice (C57Bl/6J male mice) were treated with both vehicle (0.9% Compound 4a hydrochloride (50 mg/kg, po, twice daily) for 8 weeks versus diseased control and non-diseased control LEAN-CHOW mice treated with DIO-NASH mice treated with ) was treated.

Animals were maintained and examined under the same conditions used in Example 2.2. A liver biopsy was also performed under the same conditions as in Example 2.2. A group of DIO-NASH mice were fed a high-fat diet (40% AMLN diet, D09100301 Research Diets, USA) and a control group (LEAN-CHOW) were fed a normal solid diet (Altromin 1324, Brogaarden, USA) for 35 weeks prior to testing. Denmark).

Figures 12 and 13 show treatment of DIO-NASH mice with compound 4a hydrochloride versus control DIO-NASH mice group treated with vehicle and LEAN treated with vehicle on the expression of collagen gene, a fibrosis marker. - shows the effect of the COW positive control group: collagen type I alpha 1 (COL1A1) and alpha 2 (COL1A2), type III collagen alpha 1 (COL3A1), collagen V alpha 1 (COL5A1), alpha 2 (COL5A2) and alpha 3 (COL5A3), type VI collagen alpha 1 (COL6A1), alpha 2 (COL6A2) and alpha 3 (COL6A3).

Results are given as the expression level of RPKM for each mouse group, where RPKM (reads per kilobase per million mapped reads) was calculated by normalizing total read length and number of sequencing reads to RNA A method for quantifying gene expression from sequencing data.

Data are mean±sem from 6 animals. e. m. shown as a value. ***P<0.001 vs. NASH vehicle.

Treatment of DIO-NASH model mice with the hydrochloride salt of compound 4a significantly reduced collagen gene expression compared to the control DIO-NASH mouse group. In fact, compound 4a causes regulation of more than 2000 genes, many of which are associated with NASH. Specifically, treatment of DIO-NASH mice with compound 4a (50 mg po, twice daily) for 8 weeks:
- resulted in a decrease in several proto-inflammatory markers related to monocyte recruitment, such as CD68, CCR2, MAC-2;
- resulted in the reduction of several prototypic fibrosis genes involved in astrocyte activation, such as Col1a1, Col3a1 and TIMP1;
- for inflammatory signaling, resulting in a decrease in TLR4, TGFB, and TGFBR gene expression;
- resulted in decreased expression of MAPK and AKT and increased expression of GLUT4 for insulin signaling;
- on lipid metabolism, resulted in decreased expression of CD36 and increased expression of SQLE; and - on hepatocellular cell death, resulted in increased expression of Casp7 and IL18.

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Claims (15)

of formula I for use in the prevention and/or treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), and/or symptoms and/or medical conditions associated therewith A compound, or a pharmaceutically or food acceptable salt thereof, comprising:
independently here,
R ₁ is selected from linear or cyclic mono- or dialkylamines; OR ₉ , aminoalkyl alcohols, or aminoalkyl ethers;
R ₂ is selected from benzene or heterocycle;
R ₃ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; or a benzyl group;
R ₄ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; a hydroxy or alkoxy radical; or a halogen;
_R9 is an alkyl group,
Compound. —R ₁ is NH—(CH ₂ ) _n —NH ₂ , NH—(CH ₂ ) _n —N(CH ₃ ) ₂ (where n is a value of 0 to 4); HNCH ₂ CH ₂ OH; HNCH ₂ CH ₂ OCH ₃ ; NH—N═CH-phenyl-R ₇ ; or a cyclic amine selected from:
_-R2 is selected from:
-R ₃ is selected from H, methyl, or benzyl;
-R ₄ is selected from H, methyl, methoxy, or fluorine;
-R ₇ is H or p-NO ₂ ;
—R ₈ is H, OH, or methoxy; and
-R ₅ is H, OH, or methoxy;
A compound according to claim 1 . 3. The compound of claim 1 or 2, wherein the pharmaceutically or food acceptable salt is the hydrochloride. 4a, 5a, 7a, 17a, 17b, 17c, 21a, 21b, 21c, 21d, 21e, 21f, 23a, 23b, 23c, 23d, 23e, 23f, 26a, 26b, 7b, 15a, 15b, 15c, 16a, 16b, 16c, 19a, 19b, 19c, 19d, 19e, 19f, 20a, 20b, 20c, 20d, 20e, 20f, 22a, 22b, 22c, 22d, 22e, 22f, 24a, 24b, 25a, 4. The compound of any one of claims 1-3 independently selected from or 25b.
5. The compound of any one of claims 1-4, wherein said compound is independently selected from 4a, 5a, or 7a. 6. Any one of claims 1-5, wherein the associated condition is independently selected from insulin resistance, lipid accumulation in hepatocytes, mitochondrial dysfunction, oxidative stress, apoptosis, necrosis, inflammation, or fibrosis. The compound described in 1. 6. A compound according to any one of claims 1 to 5, wherein the associated condition is cirrhosis or hepatocellular carcinoma. An effective amount for use in the prevention and/or treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) and/or symptoms and/or medical conditions associated therewith and a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient, comprising:
independently here,
R ₁ is selected from linear or cyclic mono- or dialkylamines; OR ₉ , aminoalkyl alcohols, or aminoalkyl ethers;
R ₂ is selected from benzene or heterocycle;
R ₃ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; or a benzyl group;
R ₄ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; a hydroxy or alkoxy radical; or a halogen;
_R9 is an alkyl group,
pharmaceutical composition. From insulin sensitizers, bile acid modulators, inhibitors of de novo lipogenesis, hypolipidemic agents, antioxidants, anti-inflammatory agents, immunomodulatory components, antiapoptotic agents, gut microbiome modulators, or antifibrotic agents. 9. The pharmaceutical composition of Claim 8, further comprising a second independently selected active compound. 10. A medicament according to claim 8 or 9, wherein the associated condition is independently selected from insulin resistance, lipid accumulation in hepatocytes, mitochondrial dysfunction, oxidative stress, apoptosis, necrosis, inflammation or fibrosis. Composition. 10. A pharmaceutical composition according to claim 8 or 9, wherein the associated pathology is cirrhosis or hepatocellular carcinoma. —R ₁ is NH—(CH ₂ ) _n —NH ₂ , NH—(CH ₂ ) _n —N(CH ₃ ) ₂ (where n is a value of 0 to 4); HNCH ₂ CH ₂ OH; HNCH ₂ CH ₂ OCH ₃ ; NH—N═CH-phenyl-R ₇ ; or a cyclic amine selected from:
_-R2 is selected from:
-R ₃ is selected from H, methyl, or benzyl;
-R ₄ is selected from H, methyl, methoxy, or fluorine;
-R ₇ is H or p-NO ₂ ;
—R ₈ is H, OH, or methoxy; and
-R ₅ is H, OH, or methoxy;
12. A pharmaceutical composition according to any one of claims 8-11. 13. The pharmaceutical composition according to claims 8-12, wherein the pharmaceutically acceptable salt is the hydrochloride. 14. The pharmaceutical composition according to any one of claims 8-13, wherein at least one compound of Formula I is independently selected from 4a, 5a, or 7a. At least one of Formula I for alleviating and/or preventing non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) and/or symptoms and/or medical conditions associated therewith The use of a dietary supplement comprising one compound, or a food acceptable salt thereof, and at least one food acceptable excipient:
independently here,
R ₁ is selected from linear or cyclic mono- or dialkylamines; OR ₉ , aminoalkyl alcohols, or aminoalkyl ethers;
R ₂ is selected from benzene or heterocycle;
R ₃ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; or a benzyl group;
R ₄ is selected from H; a hydrocarbon radical selected from linear or branched alkyl of 1 to 5 carbons; a hydroxy or alkoxy radical; or a halogen;
_R9 is an alkyl group,
use.